These observations give information that there are possibly other chemicals present in the MSE that could have together with NAC maintain the cell growth in media that lack nutrients thereby permitting the cells to survive longer. Tchounwou 2007) and also plays an important role in the production of glutathione to help prevent oxidative stress (De Vries and De Flora 1993). Is Kratom Illegal In Alabama Is Kratom Illegal In Alabama Counties Fifield Counties Fifield mIT (Watanabe et al 1997; Thongpradichote et al 1998) could play important roles in mediating the cytotoxicity effects seen so far. This result implies that there are
possibly other chemicals present in the leaves of this plant which could be contributor to the MSE cytotoxicity. There is an increasing popularity of use of Mitragyna speciosa Korth (Kratom) leaves as self-treatment for opioid withdrawal and chronic pain among Americans (Boyer et al 2007). This in fact reflects increasing interest in constituents of this plant MIT and its congener 7-hydroxymitragynine which have been shown to exert potent analgesic effects in various in vivo and in vitro studies (Matsumoto et al 2004). Furthermore with the recent report on the use of this plant to treat chronic pain with lesser effects of withdrawal compared to opioid prescription treatment people are using this plant as an alternative to opium drugs (Boyer et al 2008).
Similar observations were also noted for H202 MSE and MIT groups. Interestingly the majority of the cells which were treated with NAC prior to treatment with H202 appeared firmly attached to the bottom of the wells and had normal cell appearance. Brownish precipitations were also noted floating in all wells believed to be the hydrophobic fluorescent dye DCFH-DA.
Conjugation-dependant carcinogenicity and toxicity of foreign compounds Advances in Pharmacology 27: 1-512. Academic Press San Diego. ErlandssonHarris et al. High mobility group 1 protein (HMG-1) stimulates proinflammatory cytokines sysnthesis in human monocytes.
The numbers of negative wells for viability kratom legal north carolina plates and positive wells for
mutant plates were also recorded. Test Acceptance Criteria and Evaluation of the Results Following the protocols obtained from GlaxoSmithKline Company (Ware U. K) the assay was accepted based on the measurement of cytotoxicity by relative total growth (RTG) which reduced to approximately 10-20% when compared to concurrent vehicle control. The mean kratom max daily dose vehicle control value for mutant frequency (MF) are between 50-170 x 10-6 The mean cloning efficiency is between 65-120%. The mean suspension growth are between 8-32 on day 2 (following 3 hr treatment
with S9) After exclusion of obvious outliers at least 2 acceptable vehicle controls cultures remain.
M) MIT apparently stimulated cell proliferation that persisted up to 96 hr (Fig. This stimuation was small but consistent at 48 hr to 96 hr. At higher doses of MIT (3. M) cell proliferation was inhibited (Fig. These concentrations also induced substantial cell death (Fig. The IC50 of these cells at 24 hours treatment are estimated as 282.
As shown in fig. A there was a non-significant difference noted for caspase 3 and 7 activities for MSE treated cells compared to control groups at 4 hr indo white kratom powder incubation time point. For MIT treated cells (Fig.
BCA) protein assay kit from Pierce (Rockford IL). Primary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz CA) and Oncogene Research Products (Darmstadt Germany) and secondary antibodies were from Sigma-Aldrich (U. Santa Cruz Biotechnology (Santa Cruz CA).
Photo-oxidative disruption of lysosomal membranes causes apoptosis of cultured human fibroblasts. Free Radic Biol. Adulterants in herbal products can cause poisoning. kratom maeng da extract spavinaw British Medical Journal 313: 117. Long-term mutagenicity studies with chloroform and dimethylnitrosamine in female lacl transgenic B6C3F1 mice. Mutagen 31: 248-256.
SH-SY5Y cells treated with chloroform in ethanol vehicle (Fig. If chloroform contamination of MSE contributed to the toxicity of MSE then addition or synergistic cytotoxicity would be expected. M CHCl3) (Fig. This result suggests that chloroform did not enhance MSE-dependant cytotoxicity. C 5 o 1.
-5 3. D ) in MSE and MIT treated HEK 293 cells as determined by the Trypan blue exclusion assay. SH-SY5Y cells With SH-SY5Y cells low doses MSE (0. These higher doses of MSE also substantially increased cell death within 24 hr (Fig.
New apoptosis cascase mediated by lysosomal enzyme and its protection by epigallocatechin gallate. I and Mishra R. Biochemical and Biophysical Research Communications 137 813-820. Apoptosis: a basic biological phenomenon with wide ranging implications in tissue kinetics. British Journal of Cancer 26:239-257.
As shown in the table 3. MLA results for MIT in the presence or absence of rat liver S9 show no evidence of genotoxicity. The outcome of this experiment would seem to be contrary to what was seen for MSE. In the absence of rat liver S9 (Table 3.
Vehicle treated control 0. Vehicle treated control 3. D ) in MSE and MIT treated SH-SY5Y cells as determined by the Trypan blue exclusion assay.
As anticipated the experiments clearly showed that p53 was still being expressed in MIT treated groups and in control group but down regulated with time- dependant manner. M) the same pattern of p53 down regulations was seen as with the higher dose of MSE. The next experiment was carried out to further investigate if there was a correlation between p53 changes and its target gene p21 in response to MSE and kratom caps dosage MIT treatment.